Comparison of antibiotic susceptibility results obtained with Adatab* and disc methods

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1 J Clin Pathol 1984;37: Comparison of antibiotic susceptibility results obtained with Adatab* and disc methods JJS SNELL, MVS DANVERS, PS GARDNER From the Division of Microbiological Reagents and Quality Control, Central Public Health Laboratory, Colindale, London NW9 SHT SUMMARY Strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, faecal streptococci, Proteus spp, and Klebsiella spp were distributed on two occasions to two groups of laboratories, one using a commercially produced break point method (Adatab, Mast Laboratories Ltd) and the other using a disc method for susceptibility testing. Minimum inhibitory concentrations of a range of antibiotics were determined for each of the strains in the Division of Microbiological Reagents and Quality Control and a correct result of sensitive or resistant was assigned where possible to each combination of strain and antibiotic. Laboratories were asked to determine the susceptibility of the strains to those antibiotics that they would test in routine practice. Results from each laboratory were compared with the correct results. The overall error rates obtained with the Adatab and disc methods, 8% and 8-2% respectively, were not significantly different. Fewer errors were made with trimethoprim, ticarcillin, and nitrofurantoin by laboratories using Adatabs than those using discs. Fewer errors were made with gentamicin by laboratories using discs than those using Adatabs. There was no significant difference between the two groups of laboratories in reproducibility of results on repeated testing of the same strains. Laboratories using Adatabs used a wide range of different break point concentrations. The Adatab method appeared to offer no overall advantages in terms of reduced error rates or increased reproducibility of results with the strains tested. In contrast to the United States of America, where methodology could be shown, we were unable to the Kirby-Bauer method of susceptibility testing is find differences in error rates between any one of the recommended method and is widely used, there the three major methods of disc testing: Stokes' is no standard method in the United Kingdom. method, the comparative method, and Kirby-Bauer Stokes' method,' where controls are incubated on type methods.23 the same plate as the test strain, and the comparative method, where controls are on a separate plate, Adatab method by Mast Laboratories Ltd seemed Against this background, the introduction of the are the most widely used methods in the UK. Considerable variations on these basic methods have taining antibiotics (supplied by the manufacturer) interesting. In this method, tablets of a material con- been introduced by laboratories using them,2 and are dissolved in molten media, which is poured into there is no formal mechanism for updating the petri dishes. The resulting plates contain a known methods as new antibiotics are introduced. Previous concentration of antibiotic, which can be adjusted trials, in which most of the participants have used by altering the number of tablets added to a given disc methods, have shown high error rates associated with some combinations of organism and anti- then applied to the surface of the agar, usually with a volume of medium. Suspensions of organisms are biotic.23 Although in some cases association between high error rates and certain details of sensitivity is judged by growth or absence of growth multipoint inoculator, and incubated. Resistance or on one or more concentrations of antibiotic. Since Accepted for publication 23 May 1984 this method appears to be free of many of the potential problems associated with discs and is similar to * Adatab is a trade mark of Mast Laboratories Ltd. minimum inhibitory concentration (MIC) methods, 159

2 16 which are the reference method of testing, it would seem to be promising. In order to evaluate the method under field conditions a trial was organised, in which results from laboratories using break point methods were compared with those of laboratories using discs. Material and methods LABORATORIES PARTICIPATING All laboratories included in the trial were UK clinical laboratories participating in the bacteriology section of the UK National External Microbiological Quality Assessment Scheme. Laboratories were identified as using the Adatab method from questionnaires distributed as part of a previous survey.3 For each of these laboratories, a laboratory using disc methods and which was of a similar level of performance in the previous 12 month period of the bacteriology quality assessment scheme was selected as a control. This matching of performance of the two groups of laboratories was designed to ensure that differences in results between the two groups could be attributed to the method used rather than to differences in technical skill. Twenty laboratories using the Adatab method and 24 laboratories using disc methods were included in the trial, the difference in numbers between the two groups being the result of some laboratories being unable to participate. ORGANISMS Five strains each of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and faecal streptococci, all described as being from sites other than urine, and five strains each of E coli, Proteus spp, and Klebsiella spp, described as isolated from urine, were distributed. Two distributions of three cultures from each of the above groups of bacteria were made with two months between distributions. One culture in each group in the second distribution was a repeat of one included in the first distribution. The strains were selected randomly from the Division of Microbiological Reagents and Quality Control culture collection. Minimum inhibitory concentrations for these strains have been determined on several occasions in the Division, and these were repeated immediately before and during the trial by the method described previously.2 One strain of Klebsiella sp showed some contradictory results on retesting, and one strain of S aureus was later found to be coagulase negative. The results from both these strains were discarded. The MICs of the remaining strains are shown in Table 1. Snell, Danvers, Gardner ANTIBIOTICS TESTED Report forms listing for each group of organisms the appropriate set of antibiotics (as shown in Table 1) were supplied to participants, who were asked to test only those antibiotics on the list that they would normally test in routine practice. Participants were told that the trial was intended to compare the results of methods rather than to evaluate individual laboratory performance and were asked not to examine the strains more than once except in cases of gross technical failure. ASSESSMENT OF RESULTS Where possible a "correct" result of sensitive or resistant was assigned to each strain for each antibiotic tested. These "correct" results were derived by comparing the MICs of each strain with various published values for breakpoints.4 In cases where there was lack of agreement between published values the majority values were used. Correct results were not assigned where strains could not clearly be designated as sensitive or resistant to antibiotic on the basis of MIC results. Penicillinase producing strains of S aureus were designated as resistant to penicillin regardless of the MIC. Table 1 shows the classification of the strains as sensitive or resistant. The report forms used allowed the reporting of each strain by participants as sensitive, intermediate, or resistant to each antibiotic tested. Participants' results of sensitive or resistant were regarded as correct when in agreement with the designated correct result. Discrepant results were designated as follows: minor where the reported result was intermediate and the correct result was sensitive or resistant; major where the reported result was resistant and the correct result was sensitive; and very major where the reported result was sensitive and the correct result was resistant. Results NUMBER OF RESULTS RETURNED As laboratories tested only those antibiotics that they would test in routine practice, the number of reports is different for each combination of strain and antibiotic (Table 2). The difference in the number of reports received from laboratories using break points and those using discs is shown in Table 2 and reflects the fact that laboratories using discs tested a wider range of the suggested antibiotics for each organism than laboratories using Adatabs. Laboratories using Adatabs were also more restricted in the range of organisms tested-for example, some laboratories did not test urinary isolates by this method and others tested urinary isolates only.

3 Comparison of antibiotic susceptibility results obtained with Adatab and disc methods Table 1 Minimum inhibitory concentrations (MICs) ofantibiotics for the strains distributed Organism Antibiotic MICs ofstrains designated MICs of strains designated MICs ofstrains for which no as sensitive as resistant correct result was designated* Staphylococcus aureus Pseudomonas aeruginosa Klebsiella spp (from urine) Escherichia coli (from urine) Proteus spp (from urine) Escherichia coli Faecal streptococci Methicillin Tetracycline Fusidic acid Erythromycin Penicillin Amikacin Ticarcillin Tobramycin Nitrofurantoin Cephalexin Nalidixic acid Nitrofurantoin Cephalexin Nalidixic acid Cephalexin Nalidixic acid Tetracycline Cefuroxime Tetracycline Cefuroxime -5-2, , , 6-25, 6-25, , , -5-2, -5-1, -5-2, , -25-2, 1, 2-4, 1 16, 32-64, 16-32, 32-64, 32, , , 25--5, , , , 4-16, 4-16, 4-8, , 25-5, -1-1, -1-1, , 4-16, 4-8, 4-8, 4-8 8, 8-16, 8, 4, 8-16, , , , 25-5, 1, , 4-8, 8, , 2-4, 2-4, 4, 4-8, , -25-1, 5-1, -25-1, -25-1, , 2-4, , 2-4, 2-4, 4, 2-8, , -5, 5-2, 5-1, 1-2, 5 1, 2-4, 2, , 8-16, 8, , 25-5, 5-1, 1, -5-1, , 2, 2 8, 2-4, 8, 8, 8, , 1-4, 2, 1, , 1-25, 1--25, 25, 1, , 4-8, 4-8, 4-16, 4-8 4, 2-4, 2-4, 2, 2-4, , -25-1, 5-1, -25-1, 25-1, , 2-4, 2-4 2, 1, 5-2, , -5, -25-1, -5, -5, , 2 1, 1, 5-1, 1-2, 1-2, , 8-64, , 64, >128, 4-16,t 4-8,t > , ,: 16-32,: -25-4,4 8-32t 8-64, 16-64, > , 8-32, 8-32 >128, , 64-> 128, 64->128, >512, >512 > , 64->128, > >512, > , >128, >128, > > 128 >512 Where MICs differed on repeat testing, the range of MIC values obtained is shown. * Correct results were not designated where intermediate MIC values were obtained. tthese strains showed dissociated resistance to erythromycin. 4These strains were penicillinase producers. DISCREPANCY RATES The numbers of correct and incorrect results achieved by each group of laboratories are shown in Tables 2 and 3. There was no significant difference in the total error rate achieved by the two groups of laboratories (176 (8%) incorrect results and 221 > 128, 64-> 128, 64-> , >512, >512, >512, >512, >512, >512 64, , , 16-32, , 16-32, 16-32, correct results from the laboratories using Adatabs, and 382 (8-2%) incorrect results and 4271 correct results from those using discs). Significantly more very major errors were made by laboratories using Adatabs (44 (2%) very major errors and 2153 other results from laboratories using Adatabs, and 45

4 1()62 Snell, Danvers, Gardner Table 2 Comparison of the number oferrors made by laboratories using the Adatab or disc method Antibiotic Adatab Disc No of errors made No of errors made None Very Major Minor Total errors None Very Major Minor Total errors major major Staphylococcus aureus Methicillin 62 4 (6%) Tetracycline 43 1(2%) Fusidic acid 43 Erythromycin 77 () 1 (1%) Penicillin 62 8 (11%) 61 () Pseudomonas aeruginosa 77 Amikacin 15 Ticarcillin 21 Tobramycin 2 Klebsiella spp (from urine) Nitrofurantoin 19 2 (9%) 43 6 (11%) 3 8 (67%) Cephalexin 25 2 (7%) (1%) Nalidixic acid 47 9 (16%) Escherichia coli (from urine) Nitrofurantoin (2%) Cephalexin 28 2 (7%) (1 %) 1(1 %) Nalidixic acid 64 Proteus spp (from urine) 63 3 (5%) 62 3 (5%) Cephalexin 26 4 (12%) 1 (3%) (8%) Nalidixic acid 62 Escherichia coli 39 2 (4%) 5 Tetracycline () 6 (8%) Cefuroxime 56 () 4 (6%) Streptococcus faecalis 4 2 (5%) 33 5 (11%) Tetracycline 29 1 (3%) 66 8 (11%) Cefuroxime 36 2 (5%) Totals (2%) 76 (3%) 1 (1%) 5 (6%) () 1 (5%) 6 (11%) 1 (8%) 1 (4%) 1 (1%) 3 (4%) 2 (6%) 6 (9%) 3 (4%) 9 (13%) 7 (8%) 3 (14%) 13 (23%) 9 (75%) 3 (11%) 8 (11%) 9 (16%) 2 (7%) 5 (6%) 3 (5%) 4 (6%) 7 (21%) 7 (8%) 4 (9%) 6 (13%) 3 (13%) 3 (13%) 1 (1%) 3 (4%) 11 (14%) 17 (22%) 6 (1%) 9 (19%) 3 (7%) 56 (3%) 2 (5%) 14 (3%) 1 (3%) 8 (11%) 5 (12%) 176 (8%) (1 %) very major errors and 468 other results from those using discs (X2 = 12-48, p <.1)). There were apparent differences in error rates between the two groups of laboratories with the summed results for some of the antibiotics. Fewer errors were made by laboratories using Adatabs than those using discs with ticarcillin ( incorrect results and 21 correct results from laboratories using Adatabs and 21 (19%) incorrect results and 9 correct results from those using discs, (X2 = 4-72, p < -5)), nitrofurantoin (4 (5%) incorrect results and 84 correct results from laboratories using Adatabs (3%) (1%) (5%) () (2%) (9 1(1%) (1 %) (1%) 83 7 (6%) (5%) (4%) (7%) (1%) 1(1%) 2(2%) 1 (1%) 2(2%) 1 (1%) t 1(1%) 2(2%) 7 (6%) 14 (13%) 1 (23%) 1 (23%) 1 (1%) 12 (12%) 5 (24%) 1 (5%) 2 3 (3%) 3 (3%) 9 (8%) 4 (3%) 4 (3%) 3 (3%) 6(6%) 1 (%) 1 (1%) () 8 (7%) 18 (15%) 4 (3%) 3 (2%) 3 (3%) 15 (14%) 1 (1%) 4 (3%) 5 (4%) 9 (8%) 9 (8%) 4 (3%) 13 (1%) 16 (12%) 7 (5%) 3 (3%) 9 (8%) 42 (38%) 18 (16%) 4 (3%) 3 (2%) 11 (13%) 13 (3%) 27 (4%) 7 (6%) 4 (3%) 9 (7%) 1 (1%) 3 (3%) 21 (19%) 2 (47%) 15 (15%) 6 (29%) 4 (4%) 12 (11%) 8 (6%) 3 (3%) 6 (6%) 3 (2%) 26 (22%) 8 (6%) 25 (23%) 12 (9%) 7 (5%) 18 (17%) 4 (4%) 17 (14%) 23 (17%) 12 (11%) 4 (4%) 6 (55%) 4 (3%) 13 (1%) 17 (2%) 382 (8 2%) and 28 (16%) incorrect results and 144 correct results from those using discs, (X2 = 7-43, p <.1)) and trimethoprim (32 (11%) incorrect results and 254 correct results from laboratories using Adatabs and 18 (19%) incorrect results and 451 correct results from those using discs, (X2 = 9 5, p < -1)). In contrast, fewer errors were made by laboratories using discs than those using Adatabs with gentamicin (5 (1%) incorrect results and 688 correct results from laboratories using discs, and 14 (3%) incorrect results and 395 correct results from those using Adatabs, (X2 = 11-7, p < -1)).

5 Comparison of antibiotic susceptibility results obtained with Adatab and disc methods 163 Table 3 Comparison of total errors made with individual antibiotics by laboratories using the Adatab or disc method Antibiotic Method No of discrepancies Comparison of total discrepancies None Very major Major Minor Total errors Methicillin Adatab 62 4 (6%) 6 (9%) x2 6 Disc (3%) 1 (1%) 7 (6%) NS Tetracycline Adatab 93 3 (3%) 5 (5%) x2-14 Disc (<1%) 4 (1%) 18 (5%) 23 (6%) NS Fusidic acid Adatab 43 x2.1 Disc 12 4 (3%) NS Erythromycin Adatab 77 1 (1%) 3 (4%) x2 1_9 Disc 131 NS Penicillin Adatab 62 8 (11%) 1 (1%) 9 (13%) x2 2 1 Disc (5%) 9 (7%) NS Adatab (2%) 6 (1%) 14 (3%) x Disc (1%) 5 (1%) p < 1 Amikacin Adatab 15 x2-44 Disc 12 1 (1%) 3 (3%) NS Ticarcillin Adatab 21 x Disc 9 7 (6%) 14 (13%) 21 (19%) p < 5 Tobramycin Adatab 2 Disc 112 Nitrofurantoin Adatab 84 3 (3%) 1 (1%) 4 (5%) x Disc (8%) 14 (8%) 28 (16%) p < 1 Adatab (2%) 11 (4%) 15 (5%) 32 (11%) x2 9.5 Disc (<1%) 53 (9%) 53 (9%) 18 (19%) p < 1 Adatab 28 4 (2%) 14 (6%) 5 (2%) 23 (1%) x2 1 1 Disc (1%) 18 (4%) 15 (3%) 38 (8%) NS Cephalexin Adatab 79 4 (4%) 5 (5%) 3 (3%) 12 (13%) x2-66 Disc (2%) 3 (1%) 21 (7%) 31 (1%) NS Adatab (4%) 15 (4%) 15 (4%) 45 (12%) x Disc (1%) 21 (3%) 25 (4%) 55 (8%) NS Nalidixic acid Adatab (6%) 11 (6%) X2-14 Disc (1%) 14 (4%) 19 (5%) NS Cefuroxime Adatab 92 4 (4%) 5 (5%) 11 (11%) x2 X84 Disc (3%) 3 (1%) 2 (1%) 29 (14%) NS Totals Adatab (2%) 76 (3%) 56 (3%) 176 (8%) x2 8 Disc (1%) 13 (3%) 27 (4%) 382 (8X2%) NS REPRODUCIBILITY OF RESULTS results were the same on both occasions of testing One each of the strains from the seven groups of and no results were different and by the disc method bacteria was sent to participants on two separate 29 results were the same and nine results different occasions, identified on each occasion by separate on the two occasions of testing (X2 = 4-28, p < strain numbers. The results of these repeated tests -1). The ratio of the total number of results for all are shown in Table 4. Only with nitrofurantoin were antibiotics which were the same and the total there significant differences in reproducibility be- number which were different on the two occasions tween the two methods: with the Adatab method 15 of testing did not differ significantly between the two Table 4 Reproducibility on testing the seven strains distributed on two occasions Adatab No of same results on No of different results on No of same results on No ofdifferent results on each occasion of testing each occasion of testing each occasion of testing each occasion of testing Methicillin 7 2 (22%) 14 4 (22%) Tetracycline (11%) Fusidic acid 6 18 Erythromycin 11 2 Penicillin (9%) (3%) Amikacin 2 19 Ticarcillin (6%) Tobramycin 3 21 Nitrofurantoin (24%) 32 6 (16%) (25%) 29 3 (9%) 77 1 (11%) Cephalexin 1 2 (17%) 4 11 (22%) 47 3 (6%) (12%) Nalidixic acid 19 3 (14%) 5 6 (11%) Cefuroxime 11 1 (8%) 29 3 (9%) Total (8X6%) (115%) Disc

6 164 methods (267 the same and 25 (8-6%) different by the Adatab method and 674 the same and 88 (11.5%) different by the disc method, X2 = 1*97, NS). ANTIBIOTIC CONCENTRATIONS USED IN THE ADATAB METHOD The numbers of laboratories using various concentrations of antibiotics in the Adatab method are shown in Table 5. There were wide variations in the concentrations used for most antibiotics. Discussion A number of problems are inherent in this type of trial and caution is needed in interpreting the results. A particularly difficult area is the designation of "correct" results. We have, along with others, used MIC values to predict the effect of treatment, but it must be remembered that there is no absolute relation between the two. The MIC of an antibiotic for a particular organism is not an absolute value and may be influenced by such factors as inoculum density and composition of culture medium.'" A further complicating factor is the number of different break points used by various authorities4-9 and indeed by the participants using Adatabs in this trial. The MICs of the antibiotics used in this trial are in most cases unequivocal-that is, they would be interpreted as indicating resistance or susceptibility by a consensus of informed opinion. A few MICs would not be universally accepted as indicating susceptibility or resistance. In most of these cases a correct result has not been designated. In the remaining cases differences from the "correct" results may be due to differences in interpretation rather than to technical failure. Table 5 Snell, Danvers, Gardner It is unfortunate that the number of results from the control group-that is, laboratories using discs-greatly exceeded the number from laboratories using Adatabs. The purpose of the trial was to compare results from the two methods of testing in normal routine use. For this reason laboratories were presented with a list of antibiotics for each group of bacteria and asked to test only those that they would normally test in routine practice. Laboratories using discs generally tested a larger number of antibiotics for each group of bacteria and also tested all groups of bacteria. Those using Adatabs tended to test fewer antibiotics from those on the lists and also some apparently restricted the use of Adatabs to selected groups of bacteriafor example, those from urine. A further limitation of this trial was that the bacteria selected were all non-fastidious. Organisms such as Neisseria gonorrhoeae and Haemophilus influenzae would provide a different challenge to the two methods, but were not included as the selection of organisms was made from those likely to be tested routinely by both groups of laboratories. The overall error rates for the two methods (8% for Adatabs and 8*2% for discs) are comparable to the overall error rates for all methods of 5-9% and 7*1% found in two previous trials of sensitivity testing methods.23 Although there was no difference in overall error rates between the two methods, more very major errors were made by laboratories using the Adatab method. Fewer errors were made by those using Adatabs for ticarcillin, nitrofurantoin, and trimethoprim than those using discs. Fewer errors were made by those using discs for gentamicin than those using Adatabs. None of the differences in error rates with other antibiotics were significant. High error rates for trimethoprim and carbenicillin Numbers oflaboratories using various break point concentrations (pg/ml) ofantibiotics Staphylococcus aureus Methicillin 4 (8) 5 (1) 8 (2) 4,8 (1) 4,1 (1) Tetracycline 1 (3) 2 (1) 4 (1) 1,4 (3) 4,16(1) Fusidic acid -25 (1) 2 (6) 2,8 (1) Erythromycin 25 (5) 5 (3) 1 (2) 2 5 (1) -25,1 (4) -5,1 (1) Penicillin -6 (2) 25 (3) 1 (1) 2 (1) 2-5 (1) -15,-6,-25 (1) -6, 25 (2) -25,2 (3) 1 (4) 4 (3) 1,4 (1) 1,8 (3) 2,8 (2) Pseudomonas aeruginosa (2) -5,1 (1) -5,2 (1) 1,2 (1) 1,4 (5) 1,8 (4) 2,8 (3) 2,4,8 (2) Amikacin 4 (1) 4,16 (1) 8,32 (1) 16,32 (1) Ticarcillin 64 (1) 16,32 (1) 16,64 (1) 32,128 (2) Tobramycin 2 (1) -5,2 (1) 2,8 (3) Escherichia coli 8 (2) 16 (1) 24 (1) 64 (1) 8,32 (4) 5,15 (1) -5 (2) 1 (2) 4 (2) -5,2 (2) 2,8 (2) Tetracycline 1 1) 4 (2) 1,4 (1) 1,4,16 (1) 1 (1) 4 (2) 1,2 (1) 1,4 (5) 1,8 (3) 2,8 (2) 2,4,8 (1) 1 4 1) 8(31 1,87,812,321) 8,32(1) Cefuroxime 2 (1) 4 (1) 8 (2) 2,8 (2) 4,8 (2) 4,16 (1) 8,32 (3) Faecal streptococci 8 (3) 24 (1) 64 (1) 8,32 (4) 5,15 (1) -5 (3) 1 (2) 4 (2) -5,2 (2) 2,8 (1) Tetracycline 1 1) 4 (2) 1,4 (2) 1,4,16 (1) 1 (2) 4 (2) 1,2 (1) 1,8 (4) 2,8 (3) 4,1 (3) 2,4,8 (1) 2 1) 4 (1) 8 (2) 1,8 (7) 2,8 (3) 2,32 (1) 8,32 (1) Cefuroxime 2 1) 4 (2) 8 (2) 2,8 (3) 4,16 (1) 8,32 (3) Escherichia coli, Proteus spp, Klebsiella spp (from urine) Nitrofurantoin 32 (13) 16,32 (1) 4 (2) 8 (4) 16 (1) 2,8 (7) 24 (1) 32 (6) 64 (1) 1 (1) 1 (1) 8,32 (3) 32,12-8 (1) 5,15 (1) Cephalexin 32 (5) 8,16 (1) 8,32 (2) 1 (1) 4 (2) 8 (3) 1,8 (4) 2,4 (1) 2,6 (1) 2,8 (1) 2,4,8 (1) 4 (1) 8 (1) 32 (9) 1,8 (1) 2,32 (1) 8,32 (5) Nalidixic acid 16 (1) 32 (13) 16,32 (1) Numbers of laboratories in parentheses.

7 Comparison of antibiotic susceptibility results obtained with Adatab and disc methods 165 (tested instead of ticarcillin) were found in the previous two trials.23 Nitrofurantoin was not tested. ise either the disc or Adatab methods in this trial, be emphasised that no attempt was made to optim- While a reduction of these error rates is useful, it is which was designed to compare the methods as they disappointing that similar reductions were not found are performed in routine practice rather than to with methicillin, cephalosporins, sulphonamide, and evaluate the potential of the methods, which can erythromycin, all of which were associated with high best be done in a single experienced laboratory. error rates in the previous trials.23 There would appear to be little reason to recommend changes of method unless the new method can Reproducibility of test results is a factor that needs to be considered in the evaluation of any test. be shown to be more reliable, and we have not The ability to produce the same result on different shown this to be the case with the Adatab method. occasions of testing is particularly important in susceptibility testing as apparent changes in the suscep- such as ease of use, compatibility with the routine Other factors will influence the choice of method tibility of a strain may cause unnecessary changes in work, and possibilities of automation, none of which treatment of the patient. There was no significant we have evaluated in this trial. difference in the overall reproducibility between the We thank the laboratories who two methods. The number of results available for participated in this analysis of reproducibility with single antibiotics was small and it is not possible to exclude the possibility of differences between methods with individual antibiotics. As in many other areas of microbiology there is a lack of standardisation in susceptibility testing methods in the UK. Although a particular method may be widely used, variations are introduced by individual laboratories-for example, a wide range of different disc contents is used with Stokes' method.2 The Adatab method is no exception to this expression of individuality, and already a range of different concentrations of antibiotics has been introduced. Three major methods of disc testing are used: Kirby-Bauer type methods, where susceptibility is defined by strictly defined cut off points related to zone size; Stokes' method, where susceptibility is defined by comparison of test zone with the zone of a control on the same plate; and the comparative method, where sensitivity is defined by comparison of the zone size of the test strain with the zone size of a control strain on a separate plate. In contrast to the USA, where the Kirby-Bauer method is almost universally used, there is no universal method used in the UK, although Stokes' method is the single method most widely used.3 In the interests of standardisation the universal use of a single method has much to recommend it, but we have not been able to show any significant differences in error rates between these methods and have not therefore felt able to recommend the exclusive adoption of any one method.3 There are, moreover, several problems associated with disc testing" that make the use of non-disc methods such as Adatab attractive. There is no doubt, however, that it is possible to achieve a high standard of results using discs, and experience in the quality assessment scheme has shown that some laboratories consistently achieve fully correct results.23 Any method which is misused will give bad results, and this is doubtless as true of the Adatab method as it is of disc methods. It must trial for their cooperation in undertaking this extra work. References 'Stokes EJ, Waterworth PM. Antibiotic sensitivity tests by diffusion methods. ACP Broadsheet 55, Snell JJS, Brown DFJ, Gardner PS. An antibiotic susceptibility trial organised as part of the United Kingdom national microbiological quality assessment scheme. J Clin Pathol 1982;35: Snell JJS, Brown DFJ, Gardner PS. Comparison of results from two antibiotic susceptibility testing trials that formed part of the United Kingdom national external quality assessment scheme. J Clin Pathol 1984;37: Waterworth PM. Laboratory Control. In: Garrod LP, Lambert HP, O'Grady F. Antibiotic and chemotherapy. 5th ed. Edinburgh: Churchill Livingstone, 1981; Ericsson HM, Sherris JC. Antibiotic sensitivity testing. Report of an international collaborative study. Acta Pathol Microbiol Scand [B] 1971;217(suppl): NCCLS subcommittee for antimicrobial susceptibility tests. Performance standards for antimicrobial disc susceptibility tests. 2nd ed. Lancaster Avenue, Villanova, Pa 1985, USA: National Committee for Clinical Laboratory Standards, 771E, Swedish reference group for antibiotics. A revised system for antibiotic sensitivity testing. Scand J Infect Dis 1981; 13: DIN Methoden zur empfindlichkeitsprufung von bakteriellen krankheitserregern (ausser mykobakterien) geger chemotherapeutica: bewertung der minimalen hemmkonzentration. 1979: Standards Commission (Medical), German Institute for Standards, Burggrasen Strasse, 1-7, 1 Berlin 3, West Germany. 9Shannon KP, Phillips I, King BA. Aminoglycoside resistance among Enterobacteriaceae and Acinetobacter species. J Antmicrob Chemother 1978;4: ' Waterworth PM. Quantitative methods for bacterial sensitivity testing. In: Reeves DS, Phillips I, Williams JD, Wise R, eds. Laboratory methods in antimicrobial chemotherapy. Edinburgh: Churchill Livingstone, 1978;31-4. "Brown DFJ, Blowers R. Disc method of sensitivity testing and other semiquantitative methods. In: Reeves DS, Phillips I, Williams JD, Wise R, eds. Laboratory methods in antimicrobial chemotherapy. Edinburgh: Churchill Livingstone, 1978;8-3. Requests for reprints to: Dr PS Gardner, Director, Central Public Health Laboratory, Division of Microbiological Reagents and Quality Control, Central Public Health Laboratory, Colindale, London NW9 5AT, England.

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